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Journal of Clinical Microbiology, January 2003, p. 324-329, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.324-329.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Prospective Study of the Performance of Vibrational Spectroscopies for Rapid Identification of Bacterial and Fungal Pathogens Recovered from Blood Cultures

K. Maquelin,1,2 C. Kirschner,3 L.-P. Choo-Smith,1,{dagger} N. A. Ngo-Thi,3 T. van Vreeswijk,1,{ddagger} M. Stämmler,3 H. P. Endtz,2 H. A. Bruining,1 D. Naumann,3 and G. J. Puppels1*

Department of General Surgery 10M, Laboratory for Intensive Care Research and Optical Spectroscopy,1 Department of Medical Microbiology and Infectious Diseases, Erasmus MC, University Medical Center Rotterdam, 3015 GD Rotterdam, The Netherlands,2 Biophysical Structure Analyses, Robert Koch Institute, D-13353 Berlin, Germany3

Received 23 May 2002/ Returned for modification 5 August 2002/ Accepted 25 September 2002

Rapid identification of microbial pathogens reduces infection-related morbidity and mortality of hospitalized patients. Raman spectra and Fourier transform infrared (IR) spectra constitute highly specific spectroscopic fingerprints of microorganisms by which they can be identified. Little biomass is required, so that spectra of microcolonies can be obtained. A prospective clinical study was carried out in which the causative pathogens of bloodstream infections in hospitalized patients were identified. Reference libraries of Raman and IR spectra of bacterial and yeast pathogens highly prevalent in bloodstream infections were created. They were used to develop identification models based on linear discriminant analysis and artificial neural networks. These models were tested by carrying out vibrational spectroscopic identification in parallel with routine diagnostic phenotypic identification. Whereas routine identification has a typical turnaround time of 1 to 2 days, Raman and IR spectra of microcolonies were collected 6 to 8 h after microbial growth was detected by an automated blood culture system. One hundred fifteen samples were analyzed by Raman spectroscopy, of which 109 contained bacteria and 6 contained yeasts. One hundred twenty-one samples were analyzed by IR spectroscopy. Of these, 114 yielded bacteria and 7 were positive for yeasts. High identification accuracy was achieved in both the Raman (92.2%, 106 of 115) and IR (98.3%, 119 of 121) studies. Vibrational spectroscopic techniques enable simple, rapid, and accurate microbial identification. These advantages can be easily transferred to other applications in diagnostic microbiology, e.g., to accelerate identification of fastidious microorganisms.

* Corresponding author. Mailing address: Department of General Surgery 10M, Erasmus MC, University Medical Center Rotterdam, Dr. Molewaterplein 40, 3015 GD Rotterdam, The Netherlands. Phone: 31 10 4635890. Fax: 31 10 4087671. E-mail: puppels{at}hlkd.azr.nl.

{dagger} Present address: Institute for Biodiagnostics, National Research Council Canada, Winnipeg, Manitoba R3B 1Y6, Canada.

{ddagger} Present address: Department of Pediatrics, Laboratory for Immunology, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands.

Journal of Clinical Microbiology, January 2003, p. 324-329, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.324-329.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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